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GPC5基因在肺腺癌细胞系中的功能研究

, PP. 600-605

Keywords: GPC,肺腺癌,细胞增殖,细胞周期,细胞凋亡

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Abstract:

目的研究GPC5基因在肺腺癌细胞系中的表达情况及其对肺腺癌细胞系生物学行为的影响。方法采用逆转录PCR(RT-PCR)、实时定量PCR(qRT-PCR)以及Westernblot分析肺腺癌细胞系中GPC5基因mRNA及蛋白表达情况;构建GPC5过表达载体及干扰载体,通过转染构建GPC5过表达细胞系及GPC5沉默细胞系;利用CCK-8法分析肺腺癌细胞系生长情况;通过流式细胞术检测肺腺癌细胞系的周期及凋亡变化。结果检测了GPC5基因在4株肺腺癌细胞系(A549、H1299、H358和H1975)和1株肺正常上皮细胞系(HBE)中的mRNA表达情况,发现与HBE细胞系相比,GPC5基因在A549细胞系中呈低表达,在H1299细胞系中高表达,同时蛋白检测结果与mRNA水平一致;A549细胞中过表达GPC5基因可抑制细胞生长,引起细胞周期阻滞;在H1299细胞中干扰GPC5基因表达促进细胞生长,促进细胞周期;在过表达和干扰细胞模型中均未发现GPC5基因对细胞凋亡的显著影响。结论GPC5基因可以对肺腺癌细胞系的生物学行为产生影响,可能是肺腺癌的抑癌基因。

References

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